Serum and method of producing the same



Patented July 10,

. cousrmrnm LEVENTIS, or Paomux, ARIZONA.

I snnuuaun im'r non orraonncme m sum. Y

No Drawing. 7

This invention relates to antitoxinsand serums and has for its object the provision of certain improvements, in' antitoxins and Serums. More particularly, the invention 8 aims to provide improved .antitoxins and serumsand methods of producing the same.

Further, the invention is not confined to one typeor groupbut relates to all anti- 10 application to medical use and to processes of producing them. The lI1V8Ilt-10I l further aims to providenntitoxins and antibacter al Serums or antibodies in a form more'suitablefor medicaluse as a means to prevent the manifestation of the disease in the human'body organism, after infection, and protect also the body organism againstthe further offensive action of the infectious germs after manifestation of the disease, and

thus producing cure.

In 'the heretofore customary manner-of producing antitoxins and antibacterial Serums for medical use, animals are injected with specific toxins or specific infectious germs in such a manner that'the blood of the animal gradually builds up a resistanceto the specific toxin'or infectious germ with which it has been injected. After a sufiicient number" of suitable injections theblood of $0 the animal becomes soresistant that the ani-;

mal, becomes immune to that specific toxin or infectious germ; 'When the animals are thoroughly immunized they are bled and their serum is taken for medical use. A

great variety of antitoizins and antibacterial serums are made in substantially the above described manner such, for ,instance as diphtheria -antitoxin, antipneumonococcus serum and the like. j y 3' 40 Studying in general the "serotherapy against all-infectious diseases and observing the results obtained with serotherapy, I came to the conclusion, that :antitoxins and anti;

'disintoxicate the body-organism of their toxins and protect the body organism against the further offensive action of the infectiousv germs by supplying the body organism with bacterial serums do not destroy the in; fectious germs in the body organism but" protective substances.v The body organismis and check thus thefurther' Application flled January 27, 1927. smaigiwo. 104,124..

bacterial serums, against some infectious germs and mpossible to accomplish that protection against other infectious germs notably pneumonococci of all four types, I 00 came to the following conclusion:

Thebody organisms of the different ani? mals, including the human organism, react V in a diiferent Way when infected by the toxins and to all antibacterial serums,'their' same infectious germs. The common tendency is to form within the organism, and usually in theblood, protective substances against the infectious germs. Many 0I'" ganisms perform that protection easily, while others cannot succeed to protect them"- selves. The same infectious germs when in- 'troduced intoudifferent body organisms act in different manners to increase and multiply. They become more virulent or attenuated according to the body organism, culture medium, in which they find themselves. Thus, in order that two difierent body organisms, infected with the same specific germs should produce the'same protective substances they must first somehow become similar as culture media for the infectious germs. Thenguhder these conditions, antitoxins or antibacterial serums, from either of the two different body organisms, will act as protective substances when applied to the other of the two species of different body organisms which had been selected.

a As an illustration of this, if we wish to. protect a rabbit, against the infection of a specificgermfwith serum taken from a goat immunized with the same specific germ, we I must first link together the body organisms of both animals. That is, we must inject the body organism of the immunized goat with serum taken from the rabbit. Then, the body organism of the goat will not only provide a protective substances, effective, against the infectious germ, in relation to goats, as a class, but the goat body organism will at the same time provide protective substances effective against the infectious germ in relation to rabbits, as'a class. With serum thus produced,'the rabbit may be protectedagainst the infection of a specific germ with serum taken from the goat. 7 Ilhave discovered that, in a. similar manner, antitoxins and antibacterial serums may be madefrom the ,blood, of thelower animals which are effective to protect the human organism. against the infection of specifie germs. This is efiected by suitabl V I ing the immunized'lower animal human.

4 serum and then preparing the antitoxin substance or antibacterial serumfrom the of the so treated lower'animal. Thus, 1f 1t is desired. to produce humanized serum, from the donkey, effective "against pneumonia, in the human organism, the donkey,

immunized against pneumonia-,mustbe injected with human serum, a process which prior to the present invention hasn'ever been tuberculosis but without any success.

used. A serum may be thus produced which will be effective to protect. the human organism against all four types of pneumono-" cocci.

Priorto my inventlon and discovery attempts were made, with the old well known met ods, to produce polyvalent antibacterial serumagainst pneumonia'of all four types of pneumonococcusbut only with a relativesu'ccess' against pneumonia type one (1). Prior to my invention attempts were also made to prepare antibacterial serum against order that the invention may be better understood the following specific examples of the preparation of serum are given.

Preparation of poly'valent serum against pneumwnia (including all four types of pnemnonococoi).

I Based on this principle, of relation and assimilation of both interested organisms, the immunizedand the one that should be protected, I use. the following method to pro-' duce polyvalent serum, for medical use, from the donkey against human pneumonia of all four types of pneumonococci.

The animals, preferably donkeys, are previously tested withtuberculin and mallein.

The first injections are with killed pneumonococci of the four types in suspension. The first in'ections are 'made subcutaneously at intervals of six or seven days using small doses of each type administered at the same time. The doses are progressively increased until 5,000,000,000 of eac type are being injected .at a dose, avoiding in this way an severe reaction in the animals being treate The animals are then treated in the-same 'way', except that the injections are made intravenously, until a dosage of 5,000,000,000 of each type is reached. The animals'ar'e then similarly injectedintrave'nously with living n'eumonococci-of all four t pes, starting as efore with small doses an graduall increasing the size of the .dose until 5,000,000,000 of each type are being admin-. istered at each dose. This'dose is continued r until the serum ofthe animals shows a slight agglutination or precipitating power.

The animals are then permitted to rest for ten davs. 7

Following the ten day period of rest the ,animals are intravenously injected with 1 billionof living neumonococci of each type and with 5 cc. 0 human serum at the same time. The animal injected in said manner [often reacts very severely with-a high temperature, the reaction usually lastin for a few days. corresponding large ose of living pneumonococcl and human serum, if

given prior to the preliminary treatment-dc:

scribed, would kill the animal suddenly with all the symptoms of ashock of anaphylaxia.

The first injection of living germs and I man serum is followed, .at intervals of six or seven days, according to the severity'of'the' These .later injections have progressively larger reaction, with two other injections.

numbers of living germs and larger quantities of human serum, until the final dose con- -;-tains two billions of each type of the living heum'onococcus and 20 cc. of human serum.

he seventh day from the fin'alin'ection the animals are bled and the serum ta en. To each 98 cc. of theserum so produced are added, as a preservative,'2 cc. of a 2% solution of tricresol. This mixture is kept in a refrigerator for two months. The material is then tested for sterility and toxicity on guinea pigs and is then refined for medical use. This entire process of preparation re uires at least a.- five months period.

he therapeutic dose for pneumonia of all the four types-in adults is- 1 cc. for the first subcutaneous injection and the'same amount after 36 hours for the second injection. These two injections produce ve satisfactory results without any systematic reaction or interference in the efforts of thehuman organism for recovery.

Preparation of a ntituberculomlq.

The animals, preferablydonkeys, arepre viously tested withtuberculin and mallein.

I The first injectionsare with very attenuated killed tubercle bacilli in suspension and are made subcutaneously. The first doses. are

small, in the neighborhood, of 100,000000 of the very attenuated killed bacilli. rogressivel reaches five billions, avoiding in. this way any severe reaction.

Then starting with half dose, or substantially 50,000,000 of the very attenuated killed tubercle bacilli, intravethe dose is'increased until it of the former nous'injectionsare made. This dose is progressively increased in subsequent injections until the dose is increased to 5 billions. The

animal is then injected intravenously with .one. billion of the very attenuated dead tubercle bacilli in suspension together with rings on a bacilli in each field. In the place of the injection of the sputum there is formed an serum as herein described requires a period changed in any way which may seem advis' able to suit existing conditions, without de-' parting from the s irit of the invention.

abscess with a large area of inflammation. The animal becomes sick with temperature and loss of appetite. This condition continues for a week or twelve days, when the abscess is broken and inflammation and temperature subside; It takes 'two to three months for the tubercular abscess to heal completely and for the animal to increase in weight.

The animal is then allowed to rest forv another month to recover completely from the infection. Ten to twenty cc. of human serum are then injected intravenously which produces severe reaction with symptoms of anaphyla'xia- The seventh day after the in- 'ection of the human serum the animal is led and the serum taken. I

To each 98 cc. of the serum so produced are added, as a preservative, 2 cc. ofa 2%v solution of tricresol. This mixture is kept in a refrigerator for at least three months. The serum is then tested on guinea pigs for sterility and toxicity and, if found satisfactory, is refined and made ready for medical use. Other appropriate reservatives, for example, phenol or ch oroform may be employed. The-preparation of the of at least eight to ten months.

The therapeutic dose'in adults is fivedrops injected subcutaneously and is re ated-at intervals of seven days until a total of three injections have been made. After an interval of twenty days two or three more injections are made, again at intervals of seven days. The number of these second injections is governed according to-the'slight local reaction, which consists in redness and itching. The administration of the dose is accomplished without any systematic disturbance of the organism and without disturbing the welfare of the patient. After" the two preliminary series of injections,the injections are continued, as above, at intervals of two months until satisfactory results are obtained. I

While I have described certain methods by which humanized serums may be made, I do not wish to confine m selfto these specific methods, nor do I wis toconfine myself to thepreparation of the specific; substances named. The methods may -'be varied or While I have described methods in which dead, attenuated or living bacteria were used, I do not wish-to thus confine myself,

but, may instead,'make use of toxins'given ofi by the living bacteria or liqui'd'extracts from .cultures, disintegrated micro-organisms or disintegrated products of the bacterial any way with the production of the specific affection.

' I claim:

1. In the process of preparing antitoxins and'antibacterial serums, treatin an animal with an antigen, in'ecting serum rom an ammal to be protecte and of a different species into. said treated animal and withdrawing the serum from the treated animal.

2. In the process of preparing antitoxins and antibacterial serums for specific animal use, injecting an animal of a species other thamthe specific animal with the toxin or bacterial extract, injecting the animal with the specific animal serum and withdrawing the serum from thejtreated animal.

3. In the process of preparin antitoxins and antibacterial serums for man use, treating an animal with an antigen, injectin human serum into the said treated anima and withdrawing serum from the thus treated animal.

4. In the process of preparing antitoxins and antibacterial 'serumsfor human use, injecting an animal with a specific germ, injecting human serum into said treated animal, withdrawing serum from the thus treated animal, and preparing the withdrawn serum for medical'use.

animal against an in ectious germ specific to the first animal by the injection into the animal organism of specific animal serum which makes the second animal-organism more su's- 5. In the process of preparing polyyalent ceptible to the said specific infectious germ. t

. i; In the process of preparing antitoxins and antibacterial serums, treating an animal not normally responsive to'a specific disease with thcspecific' antigen, injecting into the said treated animal serum from an animal normally responsive to the specific diseasethereby making the said treated animal res onsive to :--the specific disease, and withrawing serumfrom the thus treated animal. .8. A specificserum derived from the blood of an animal which is not normally responsive to the specific disease, but which has animal. Y

In' testimony'whereof I aflix my signature. I coNSTANTINE LEVENTIS. M. D.

beeuf'made responsive by injection with the specific antigen and serum .of a responsive 

